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Critical residues involved in tau binding to fyn: implications for tau phosphorylation in Alzheimer's disease
Dawn H. W. Lau1,
Marte Hogseth1,
Emma C. Phillips1,
Michael J. O'Neill2,
Amy M. Pooler1,3,
Wendy Noble1 &
Diane P. Hanger1
Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterised by neuropathological deposits of amyloid plaques and neurofibrillary tangles comprised of β-amyloid and tau protein, respectively. In AD, tau becomes abnormally phosphorylated and aggregates to form intracellular deposits. However, the mechanisms by which tau exerts neurotoxicity in disease remain unclear. Recent studies have suggested that the presence of tau at synapses may indicate a role in neuronal signalling, which could be disrupted in pathological conditions. The non-receptor-associated tyrosine kinase fyn is located at the dendrite in neurons, where it was recently shown to interact with tau to stabilise receptor complexes at the post-synaptic density. Fyn also co-localises with tau in a proportion of neurons containing tau tangles in AD and fyn is also a tau kinase. Hence, tau-fyn interactions could play a pathogenic role in AD. Here we report the identification of critical proline residues, Pro213, Pro216, and Pro219, located within the fifth and sixth Pro-X-X-Pro motifs in the proline-rich region of tau, that are important for its binding to fyn. These residues in tau are flanked by numerous phosphorylation sites and therefore we investigated the relationship between fyn and the degree of tau phosphorylation in human post-mortem brain tissue. We found no difference in the amount of fyn present in control and AD brain. Notably, however, there was a significant correlation between fyn and phosphorylated tau at specific phospho-epitopes in control, but not in AD brain. Our results suggest that the pathological mechanisms underlying AD, that result in increased tau phosphorylation, may disrupt the physiological relationship between tau phosphorylation and fyn.
The microtubule-associated protein tau has long been recognised as having a role in stabilising microtubules in neurons, particularly in axons. Recent evidence however, suggests that tau is also present in dendrites where it may play a role in synaptic function [1–3]. In support of this suggestion, tau has been shown to associate with synaptic proteins, such as post-synaptic density protein 95 (PSD-95) and fyn [1, 4]. In particular, a functional role has been shown for the interaction between tau and fyn, whereby tau directs fyn to the post-synaptic density to interact with N-methyl-D-aspartate (NMDA) receptors [1]. Direct binding of endogenous tau and fyn was first shown in SH-SY5Y neuroblastoma cells [5]. Subsequently, a detailed study in mouse brain demonstrated that a small proportion of tau interacts with fyn in dendrites [1]. Using a truncated form of tau lacking the microtubule-binding domain that binds to fyn but is unable to associate with microtubules, it was shown that tau sequesters fyn, thereby excluding it from dendrites and causing fyn to be translocated into the neuronal cell body [1]. Furthermore, expression of this truncated form of tau destabilised the interaction between fyn, NMDA receptors, and PSD-95 [1]. Crossing amyloid precursor protein (APP) mutant mice with transgenic mice expressing truncated tau ameliorated the premature mortality and memory deficits, and reduced the susceptibility to excitotoxic seizures observed in APP transgenic mice [1]. These data indicate that the mis-localisation of fyn caused by truncated tau rescued amyloid-induced phenotypes. Therefore, tau-fyn interactions could play a functional role in Alzheimer's disease (AD). We have previously demonstrated a role for fyn in the dynamic targeting of tau to the plasma membrane in neurons [6]. We found that the phosphorylation status of tau affects its trafficking between the cytosol and the membrane in neurons, and that this was dependent on the region of tau that is phosphorylated [6]. Thus there is mounting evidence that tau-fyn interactions could be involved in new functionalities for tau at dendrites and in synaptic signalling [7, 8].
We focused here on identifying the binding site(s) for the tau-fyn interaction. It is well known that SH3 domains of src family kinases, of which fyn is a member, bind to PXXP motifs [9]. Tau has seven such motifs, some overlapping in tandem, within the proline-rich region in the N-terminal half. Therefore it is of interest to determine which of the motifs is critical for tau-fyn interactions. Previous studies to identify the critical PXXP motif have suggested that the sixth and seventh PXXP motifs (P216–P219 and P233–P236, respectively) are important for tau-fyn binding [10, 11], while other studies suggested that the seventh PXXP motif, rather than the sixth PXXP motif, was more important for tau-fyn binding [1, 5].
In this study, we extend the findings from Lee et al. [5] and Usardi et al. [10] to include investigation of the remaining PXXP motifs on tau using full-length, intact tau to avoid structural changes. Furthermore, since phosphorylation at serine and threonine residues affects interactions between tau and fyn [7] and the relationship between tau and fyn has not been explored extensively in humans, we investigated whether phosphorylation of tau around these motifs would correlate with levels of fyn in human post-mortem brain tissue from unaffected individuals or patients with AD.
A plasmid expressing the longest isoform of human CNS tau containing two N-terminal inserts and four microtubule-binding repeats (2N4R) has been described previously [12]. The generation of plasmids encoding 2N4R tau containing proline-to-alanine substitutions at residues 216 (P216A) or 233 (P233A) of tau has been described previously [10]. Site-directed mutagenesis (QuikChange II XL, Agilent Technologies) was used to produce 2N4R tau constructs harbouring individual proline-to-alanine substitutions, P176A, P182A, P200A, P206A, P213A, and P219A. The primers used are shown in Table 1. Wild-type (WT) and mutant tau cDNA were subcloned into the pcDNA3.1-V5-His-TOPO vector (Invitrogen) to generate plasmids expressing V5-His-tagged tau. Plasmids encoding glutathione-S-transferase (GST), or GST fused to the N-terminus of the src homology (SH) two or three domains of fyn were a gift from Dr S. Anderson (University of Colorado Anschutz Medical Campus, Aurora, CO, USA) [13].
Table 1 Primers used for site-directed mutagenesis
Cell culture and transfection
Chinese hamster ovary (CHO) cells were maintained in Ham's F-12 (GE Healthcare) supplemented with 10 % (v/v) foetal bovine serum (Biosera), 2 mM L-glutamine, 100 U/ml penicillin, and 100 μg/ml streptomycin, and incubated in a 5 % CO2 atmosphere at 37 °C. Cells were plated at a density of 200,000 per well of a 6-well plate and plasmids were transfected 24 h after plating using jetPEI® (Polyplus Transfection), according to the manufacturer's instructions. Cells were harvested 24 h after transfection.
Pull-downs with glutathione S-transferase fusion proteins
Frozen glycerol preparations of E.coli containing GST- and GST-fyn-SH3-encoding plasmids were streaked across agar plates and incubated overnight at 37 °C. A single bacterial colony was used to inoculate 5 ml LB-amp broth and incubated overnight at 37 °C with shaking, before further inoculating 250 ml LB-amp broth and incubating for two hours at 37 °C with shaking until an optical density of 0.6–1.0 nm was reached. 1 mM isopropyl-1-thio-β-D-galactopyranoside (IPTG) was added to the cultures and incubated for a further 2 h with shaking. The culture was then centrifuged at 20,000 g(av) for 15 min at 4 °C to pellet the bacteria. The pellet was resuspended in 12 ml TNE buffer (25 mM Tris-HCl, pH 7.5, 100 mM NaCl, 1 mM ethylenediaminetetraacetic acid (EDTA), and complete protease inhibitor cocktail without EDTA [Roche]) and homogenised by sonication (Vibra-Cell™, Sonics & Materials Inc.) for six 15 s bursts on ice. Triton X-100 was added to a final concentration of 1 % (v/v), mixed, and the samples were centrifuged at 30,000 g(av) for 10 min at 4 °C. The supernatant, containing GST-fusion proteins, was collected and stored on ice. A 50 % (v/v) slurry of glutathione-4B beads (GE Healthcare, Buckinghamshire, UK) was washed three times in TNE buffer and added to supernatant containing GST-fusion proteins and rotated for 90 min at 4 °C, then centrifuged at 3300 g(av) for 1 min at 4 °C to pellet the beads. Beads coupled to GST-fusion proteins were washed three times with TNE buffer and resuspended as a 50 % (v/v) slurry.
CHO cells transfected with WT or mutant tau were washed with phosphate-buffered saline, harvested into ice-cold lysis buffer (25 mM Tris-HCl, pH 7.5, 10 % (v/v) glycerol, 0.5 % (v/v) Triton X-100, 1 mM EDTA, 1 mM ethylene glycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid, 150 mM NaCl, complete protease inhibitor cocktail without EDTA [Roche], and phosphatase inhibitor cocktail 2 [Sigma]), and incubated on ice for 15 min. Lysates were centrifuged at 6000 g(av) for 5 min to pellet cell debris. GST-beads (50 % slurry) were washed twice in wash buffer (50 mM Tris-HCl, pH 7.5, 2 mM EDTA, 0.5 % (v/v) Triton X-100, 150 mM NaCl, complete protease inhibitor cocktail without EDTA [Roche], and phosphatase inhibitor cocktail 2 [Sigma]) and incubated with the cell lysates. The pre-cleared cell lysates were incubated overnight at 4 °C with washed GST-beads or GST-fyn-SH3-beads. Beads were pelleted at 6000 g(av) for 5 min and the supernatants were discarded. Beads were washed twice with wash buffer, then Laemmli sample buffer was added and the samples heated at 100 °C for 5 min to elute bound protein. Proteins were separated on 10 % (w/v) sodium dodecyl sulphate (SDS)-polyacrylamide gels, transferred to nitrocellulose membranes and western blots were probed with antibodies to total tau, as described below. The amount of tau bound to GST-fyn-SH3 beads was quantified using an Odyssey scanner (Li-Cor Biosciences) and standardised against the total amount of tau in the corresponding cell lysate.
Post-mortem human brain tissue
Samples of frozen temporal cortex from post-mortem human brain (15 AD and 11 controls) were obtained from the London Neurodegenerative Diseases Brain Bank at the Institute of Psychiatry, Psychology and Neuroscience, King's College London (details in Table 2). Samples were collected and distributed in accordance with local and national research ethics committee approvals (REC reference: 08/MRE09/38+5). Alzheimer disease-related pathology was classified using diagnostic neuropathological criteria from the consortium to establish a registry for Alzheimer's disease (CERAD) and Braak staging [14, 15]. The average ages of the AD and control cases were 87.7 ± 1.4 years and 79.2 ± 3.6 years (mean ± s.e.m), respectively. The average post-mortem delays for the AD and control brains were 33.5 ± 5.5 h and 24.2 ± 4.8 h (mean ± s.e.m), respectively. Brain tissue was thawed on ice and homogenised at 100 mg/ml in 2× Laemmli sample buffer using a mechanical homogeniser (Ultra Turrax® T8, Werke GmbH & Co., Germany). Homogenates were briefly sonicated (Vibra-Cell™, Sonics & Materials Inc.) before being centrifuged at 16,000 g(av) at 4 °C for 20 min. The supernatants were collected and stored at −80 °C.
Table 2 Case details for control and late-stage Alzheimer's disease post-mortem human temporal cortex
Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and western blots
Proteins were separated on 10 % (w/v) SDS-polyacrylamide gels and transferred to nitrocellulose membranes. After blocking, membranes were incubated overnight at 4 °C with primary antibodies directed against total tau (rabbit polyclonal, 1/10,000, DAKO; mouse monoclonal CP27, 1/10,000, gift from Peter Davies), GST (goat polyclonal, 1/2000 GE Healthcare), phosphorylated tau (mouse monoclonal PHF-1, 1/5000, and CP13, 1/400, both gifts from Peter Davies; rabbit polyclonal pS214, 1/500, Abcam; rabbit polyclonal pS262, 1/500, Abcam), dephosphorylated tau (mouse monoclonal Tau-1, 1/5000, Millipore), fyn (rabbit polyclonal, 1/1000, HPA023887, Sigma), β-actin (mouse monoclonal, AC15, 1/10,000, Sigma), and neuron-specific enolase (mouse monoclonal, BBS/NC/VI-H14, 1/10,000, DAKO). Blotted membranes were incubated with secondary antibodies (Alexa Fluor® 680 goat anti-mouse, 1/10,000, Life Technologies or IRDye™ 800 conjugated goat anti-rabbit, 1/10,000, Rockland Immunochemicals Inc.) and antigens were visualised and quantified using an Odyssey scanner (Li-Cor Biosciences).
Colloidal Coomassie blue staining of polyacrylamide gels
For analysis of GST proteins bound to glutathione beads, SDS-polyacrylamide gels were incubated in ProtoBlue Safe Working Solution (National Diagnostics) and protein bands were imaged using an Odyssey scanner (Li-Cor Biosciences).
Data were tested for normal distribution using the D'Agostino-Pearson normality test. Normally distributed samples were analysed for statistical significance (p < 0.05) using Student's t-test or ANOVA. Data that were not normally distributed were analysed using Mann-Whitney or Kruskal-Wallis tests. Pearson's or Spearman's rank correlation was used to determine correlations between proteins, as appropriate. GraphPad Prism five was used for all statistical analyses.
Interaction between the fyn-SH3 domain and tau is modulated by PXXP motifs in tau
To determine which PXXP motif is important for binding of tau to fyn-SH3, we used a GST pull-down assay to test fyn-SH3 binding with WT tau and eight mutant PXXP tau constructs (P176A, P182A, P200A, P206A, P213A, P216A, P219A, and P233A) in which individual proline residues were substituted by alanine (Fig. 1). GST fusion proteins (GST-only or GST-fyn-SH3) coupled to glutathione beads were prepared and the purity of the GST proteins bound to the beads was assessed on Coomassie blue-stained polyacrylamide gels. Following induction of protein expression with IPTG, GST or GST-fyn-SH3 was readily observed in the bacterial cell lysate (Fig. 2a). Analysis of bound proteins showed single bands at the predicted sizes for GST (27 kDa) or GST-fyn-SH3 (32 kDa).
PXXP motifs in the proline-rich region of full-length human tau. The amino acid sequence of the longest isoform of human CNS tau is shown, with the PXXP motifs underlined and numbered (1–7). Individual proline to alanine substitutions within the PXXP motifs are indicated by asterisks
Identification of PXXP motifs critical for tau binding to fyn-SH3. a Coomassie blue-stained gels of supernatants from cleared bacterial lysates following induction of protein expression (Binding S/N), show glutathione S transferase (GST) and GST-fyn-SH3 at the expected sizes of 27 and 36 kDa, respectively. Specific coupling of GST or GST-fyn-SH3 proteins is shown (Beads). Molecular weight markers (kDa) are shown on the left. b Western blots of GST-fyn-SH3 pull-downs from CHO cells expressing wild-type (WT) or mutant P → A tau probed with antibodies recognising tau (upper panel) or GST (middle panel). CHO cell lysates were probed with tau antibody for standardisation (lower panel). Lanes spliced from the same blot are indicated by a dotted line. Molecular weight markers (kDa) are shown on the left. c Quantification of the amount of tau immunoreactivity pulled down by GST-fyn-SH3, as a proportion of WT tau, following standardisation to total tau in the corresponding lysate. Values represent mean ± SEM, n = 6–22. One-way ANOVA with post-hoc Dunnett's test, **p < 0.01, ***p < 0.001
CHO cells were transfected with WT or mutant PXXP tau and cell lysates were incubated with GST or GST-fyn-SH3 beads. Proteins bound to the beads were analysed on western blots probed with an antibody against total tau (Fig. 2b). The amount of tau bound to fyn-SH3 beads was quantified and standardised against the total amount of tau in each cell lysate (Fig. 2c). Western blots of the fyn-SH3 beads showed that the amounts of P216A and P219A tau bound to fyn-SH3 was significantly decreased to ~65 % of SH3-bound WT tau (Fig. 2b, c; p < 0.01). Interestingly, the amount of P213A tau bound to fyn-SH3 was significantly increased by 50 % compared to WT tau (Fig. 2b, c; p < 0.001). There were no significant differences between the amounts of any of the other PXXP tau mutants tested (P176A, P182A, P200A, P206A, or P233A), compared to WT tau bound to fyn-SH3 (Fig. 2b, c). These results indicate that the PXXP motif at P216-P219 of tau is important for maintaining interactions between tau and fyn-SH3. Furthermore, our findings show that disruption of a proline residue closely apposed to the N-terminus of the sixth PXXP motif also increases the interaction of tau with fyn-SH3.
In agreement with previous studies reported by our group and others [10, 11], binding of tau to fyn-SH3 was unaffected by substitution of P233 with alanine in the seventh PXXP motif in tau (Fig. 2b, c). Despite reports that removal of the PXXP motif at 233–236 of tau inhibits its binding to fyn-SH3 [5], our data supports the idea that PXXP motifs N-terminal to P233 in tau are more important for tau-fyn binding. However, it should be noted that whilst both the individual P216A and P219A tau mutations reduced tau binding to fyn-SH3 by approximately 35 %, neither mutation completely abolished the interaction of tau with fyn-SH3. Thus it is likely that there are other regions of tau, in addition to the individual proline residues located inside the PXXP motifs, which are important for tau binding to fyn-SH3.
Tau phosphorylation in Alzheimer's disease brain is unchanged at sites surrounding critical PXXP motifs
We and others have shown previously that tau phosphorylation can reduce its binding to fyn-SH3 [6, 7, 16]. The PXXP motif at residues 216–219 resides within a region of tau that is enriched in serine/threonine phosphorylation sites [17]. Thus abnormal phosphorylation of tau in AD could result in altered interactions between tau and fyn-SH3. To determine the relationship between tau phosphorylation and fyn, post-mortem human brain tissue from controls (unaffected individuals and Braak stages I–II) and from individuals with moderate to late stage AD (Braak stages IV–VI) at autopsy was analysed by western blot [18]. Probing with an antibody against tau showed that, following standardisation to neuron-specific enolase (NSE), the total amount of tau is highly variable in post-mortem human brain, particularly in AD (Fig. 3a). NSE standardisation allows for variable neuronal loss due to differing degrees of neurodegeneration in the human brain samples. The total amount of tau present in AD brain at Braak IV–VI stages was significantly increased compared to control brain (Fig. 3a; p < 0.01), in agreement with previous reports [19, 20]. Probing the blots with an antibody against fyn showed that after standardising to NSE, fyn was more consistent within both control and AD brain, and that the total amount of fyn was not significantly different between control and AD brain (Fig. 3b).
Total tau is increased in AD brain whilst the amount of fyn is unchanged. a Western blots of control and Alzheimer's disease (AD) post-mortem human brain tissue probed with antibodies to total tau (upper panel) and neuron-specific enolase (NSE, lower panel). The amount of tau was quantified relative to NSE in each sample (right). b Western blots of control and AD brain tissue probed with antibody against fyn (upper panel). The amount of fyn was quantified relative to NSE in each sample (right). Molecular weight markers (kDa) are shown on the left. Values represent mean ± SEM, n = 11–15 per group. Mann-Whitney non-parametric test, **p < 0.01, NS = not significant
Next, we analysed the human post-mortem brain tissue for tau phosphorylation, particularly at serine/threonine residues lying in close proximity to the proline-rich region, and also towards the C-terminus of tau. Tau phosphorylation was standardised to total tau, to take into account the wide variation in the amount of tau present in each brain sample. Tau-1, an antibody which recognises tau when it is dephosphorylated at S199/S202, revealed prominent bands corresponding to normally phosphorylated tau in all samples tested (Fig. 4a). Quantification of Tau-1 showed a 40 % decrease in Tau-1 immunoreactivity in AD compared to control brain, indicating increased phosphorylation of S199/S202 tau in AD, as expected [21] (Fig. 4a; p < 0.01). Tau immunoreactivity to CP13 antibody (tau phosphorylated at S202) was barely detectable in controls but immunoreactive bands at ~65–70 kDa were recognised by this antibody in AD brain (Fig. 4b), as shown previously [22]. The Odyssey imaging system has a dynamic range of several orders of magnitude and is capable of detecting immunoreactive bands that may not be visible by eye, as demonstrated by the presence of CP13-immunoreactive bands in control brain tissue in high-intensity scans (Additional file 1: Figure S1a). Quantification of these CP13-immunoreactive tau species relative to total tau revealed that phosphorylation at S202 of tau is significantly increased in AD brain compared to controls (Fig. 4b; p < 0.01). Quantification of blots probed for phosphorylation of tau at S214, located within the fifth PXXP motif, showed that this site on tau is phosphorylated similarly in both AD and control brain, although the degree of phosphorylation showed a larger degree of variation in control brain (Fig. 4c). Similarly to CP13, the pS262 tau antibody detected 65–70 kDa species in AD brain but not in controls and these tau species were also found to be significantly increased in AD, as previously shown [23] (Fig. 4d; p < 0.01). The PHF-1 antibody (tau phosphorylated at S396/S404) showed barely any immunoreactivity in control brain and a significant increase in AD brains compared to controls (Fig. 4e; p < 0.05), although weak PHF-1 immunoreactivity can be observed in control brain tissue in high intensity blots (Additional file 1: Figure S1b). Taken together, these data show that consistent with previous reports, increased phosphorylation at tau epitopes corresponding to S199, S202, S262, and S396/S404 are a feature of AD brain [20, 24–29].
Altered tau and fyn expression at late stages of AD. Western blots of control and Alzheimer's disease (AD) post-mortem human brain tissue probed with antibodies recognising (a) tau dephosphosphorylated at S199/S202 (Tau-1), b tau phosphorylated at S202 (CP13), c tau phosphorylated at S214, d tau phosphorylated at S262, and (e) tau phosphorylated at S306/S404 (PHF-1). Quantification of each epitope relative to total tau is shown on the right. Molecular weight markers (kDa) are shown on the left. Values represent mean ± SEM, n = 11–15 per group. Mann-Whitney non-parametric test, *p < 0.05, **p < 0.01, NS = not significant
Fyn is not correlated with tau phosphorylation in Alzheimer's disease brain
To determine whether there is a correlation between the amount of fyn protein in AD and tau phosphorylation at the epitopes investigated in this study, we performed a non-parametric Spearman, or parametric Pearson correlation analysis, depending on the normality of the data distribution. Correlation between tau and fyn was first examined in the combined cohort of control and AD patients, to investigate whether there was any association irrespective of neuropathological status. We found no significant correlation between the amount of fyn and either total tau, or tau phosphorylated at S199/S202, S202, S214, S262, or S396/S404 in the combined samples (data not shown).
We next determined whether there was a correlation between fyn and tau in the separate control and AD cohorts that may be due to their differential neuropathological AD status (Fig. 5). A summary of the correlation coefficients and significance levels obtained for correlation between fyn and tau in control and AD brain is shown in Table 3. This analysis revealed the interesting findings that the amount of fyn present in control human brain correlated positively with the amount of tau phosphorylated at S202 (CP13), S262, and S396/404 (PHF-1), but not at S214 (Table 3). In contrast there was no correlation between fyn and phosphorylated tau in AD brain, suggesting a potential disruption in the fyn-tau relationship in AD. Our data support the hypothesis that there may be an association between tau and fyn under physiological conditions, but that pathological activation of fyn or other disease-associated alterations in fyn in AD may directly or indirectly impact regulation of tau phosphorylation, or vice versa.
Correlation analysis of disease-related changes in tau and fyn in Alzheimer's disease brain. a Total tau, expressed relative to neuron-specific enolase (NSE), and (b-f) tau phospho-epitopes Tau-1; CP13; pS214; pS262; and PHF-1, expressed relative to total tau were compared to the amount of fyn relative to NSE in post-mortem control and Alzheimer's disease (AD) human brain. Pearson's or Spearman's rank correlation analysis showed significant correlation of fyn with CP13, pS262, and PHF-1 in control brain, but not in AD brain (details in Table 3), as indicated by a line of best fit
Table 3 Correlation analysis between tau and fyn in control and Alzheimer's disease brain
The interaction between tau and fyn has recently been proposed as a possible therapeutic target for AD [11, 30–32]. Aberrant mislocalisation, or post-translational modification, of tau and/or fyn under pathological conditions could lead to altered binding between these two proteins [33]. A growing body of evidence supports the hypothesis that tau-fyn interactions play a role in regulating synaptic function [1, 8, 34]. Altered binding between tau and fyn could lead to unwanted downstream consequences in AD, triggering further cascades that lead to synaptotoxicity. In particular, tau and fyn appear to work in concert to promote NMDA receptor stability [1, 34]. Furthermore, inhibiting fyn kinase activity reduces tau aggregation in a transgenic mouse model of AD, suggesting that tau-fyn interaction could exacerbate tau pathology in AD [30, 35]. Therefore, we sought to identify the physiologically important binding site between tau and fyn, and to determine whether disease-related phosphorylation of tau was related to levels of fyn in pathogenic conditions.
In this study, we substituted individual proline residues with alanine in each of the seven PXXP motifs in tau to determine the effect on its ability to bind to the SH3 domain of fyn. We found that substitution of either P216 or P219 in tau reduces tau binding, whereas the P213A mutation increases the association of tau with fyn-SH3. This indicates that fyn-SH3 binds to the sixth PXXP motif of tau (residues 216–219), and that the fifth PXXP motif of tau at 213–216 also plays an important role in regulating tau-fyn-SH3 interaction.
We have demonstrated previously that replacing P216, but not P233, with alanine reduces tau binding to fyn-SH3 in CHO cells [10], and this finding has since been confirmed in a more recent study using bioluminescence resonance energy transfer in cells [11]. In contrast, earlier in vitro studies, using truncated tau constructs, have suggested that the seventh PXXP motif at residues 233–236, rather than the sixth motif (216–219) is more important for tau-fyn binding [1, 5]. The reason for this discrepancy is not known, but one possible explanation could be differential binding and conformation of full-length, intact human tau that was used in this study and others that identified the sixth motif (P216–P219) [10, 11]. In comparison, truncated tau was used in other studies that identified the seventh PXXP motif (P233–P236) [1, 5].
PXXP-containing ligands are categorised as either class I or II, with consensus sequences of RPL[PPXP] or XXX[PXXP]XR respectively, depending on whether the flanking by positively charged residues are located N or C terminal to the PXXP motif [36]. Class I and II ligands differ in the orientation in which they bind to SH3 domains and, whereas SH3 domains can exhibit a preference for either class of ligand, fyn-SH3 binds to both classes [36–38]. In contrast to previous reports in which the tau PXXP motif at 233–236 appeared to be important for binding to fyn-SH3 [1, 5], we show here that tau residues 213–219 are responsible for this interaction, with the sixth motif (P216–P219) enabling binding. Furthermore, we have identified a potential regulatory role for P213 in the fifth PXXP motif in tau since mutation of this residue results in increased binding to fyn. The amino acid sequences flanking the fifth, sixth and seventh PXXP motifs in tau are 208-SRSRT[PSLP]TPPT-220, 211-RTPSL[PTPP]TREP-223, and 228-VVRTP[PKSP]SSAK-240, respectively. Therefore, the fifth (P213–P216) and seventh (P233–P236) PXXP motifs show some similarity with class I ligands, whereas the sixth motif (P216–P219) meets the consensus requirements of a class II ligand. Although fyn-SH3 can bind to both class I and II ligands, interaction with tau could be dependent on its conformation, such that fyn-SH3 might favour binding to the sixth PXXP motif as a class II ligand. However, this has not yet been examined and the functional implications of SH3 domain binding to class I or II PXXP ligands are not well defined.
The sixth tau PXXP motif (P216–P219) appears to be the preferred binding domain for fyn-SH3, although disruption to this region of tau could enable fyn to bind to alternative PXXP motifs in tau. The fact that none of the individual proline to alanine substitutions in tau examined here completely abolished fyn-SH3 binding, suggests that fyn-SH3 has the capacity to bind to different PXXP motifs under different physiological and pathological conditions. Further support for this hypothesis comes from a study by Ittner et al. [1] who demonstrated that a truncated tau construct (residues 1–196), containing the N-terminus and the first two PXXP motifs, did not bind fyn in transfected HEK293T cells, whereas another truncated N-terminal tau construct (1–221), that included the sixth PXXP motif, exhibited reduced binding to fyn compared to full-length tau. Thus, exclusion of the seventh PXXP motif (residues P233–P236) and the remaining C-terminal residues was insufficient to completely abolish tau-fyn-SH3 binding [1], suggesting a role for other regions of tau in mediating binding with fyn-SH3.
Interestingly, we found that the P213A tau mutation resulted in an increased amount of tau bound to fyn-SH3. One possibility is that the alanine substitution at this residue alters the conformation of tau to make it more favourable for binding to fyn-SH3, providing that the sixth PXXP motif is also present. It has been shown that specific residues outside the core PXXP ligand can contribute to the binding between PXXP motifs and SH3 domains [39], which could explain the increased binding affinity between P213A mutant tau and fyn-SH3. For example, a serine residue located close to the PXXP motif is important for binding to the SH3 domain of the adaptor protein, Nck (non-catalytic region of tyrosine kinase adaptor protein 1) [40]. Residues located C-terminal to PXXP motifs can also modulate binding to fyn-SH3, although residues N-terminal to PXXP have not yet been examined [41]. Thus, if fyn-SH3 binds to tau at residues 216–219, the proline to alanine substitution at 213 of tau could result in increased binding affinity with fyn-SH3. Another possibility is that the P213A mutation may alter tau phosphorylation around this PXXP motif, which could enhance tau-fyn binding at the SH3 domain.
In a related study, we showed previously that the phosphorylation status of residues S199/S202 in tau influences its binding to fyn-SH3 [6], but the effects of tau phosphorylation at other residues on tau-fyn binding have not been determined. The proline-rich region of tau, which contains the PXXP motifs, also includes a high density of serine/threonine residues [17]. Therefore, it is likely that phosphorylation, or other post-translational modifications, of residues surrounding the sixth PXXP motif of tau, could influence its binding to fyn-SH3. Substitution of 18 serine and threonine residues in the N-terminal half of tau to glutamate, to mimic a state of permanent phosphorylation, inhibits its binding to fyn-SH3 [7]. These mutations include 11 residues which are located within the proline-rich region of tau. Binding of tau to fyn-SH3 was also abolished using Tau-Glu10, a construct that mimics permanent phosphorylation at ten serine/threonine sites, including five in the proline-rich region [7]. Additionally, fyn-SH3 is unable to bind to highly phosphorylated tau extracted from AD brain [7]. Therefore, it appears that serine/threonine phosphorylation of tau influences its interaction with fyn-SH3, and much of this phosphorylation occurs around the PXXP motifs (P213-P219) identified here as being important for binding between tau and fyn-SH3.
However, the relationship of fyn with total tau or tau phosphorylation has not been previously investigated in AD. In agreement with previous reports, and despite the wide variation between individuals with AD, we detected increased tau in AD brain [19, 20].
We found no significant correlation between the amount of fyn present and the degree of tau phosphorylation in AD brain tissue, suggesting that modification of tau phosphorylation, at least at the serine/threonine residues examined in this study, does not result or lead to increased fyn in AD. Interestingly, in control brain tissue, although the amount of fyn did not correlate with tau phosphorylation at residues S199 or S214, there was a significant positive correlation between fyn and tau phosphorylated at S202, S262, and S396/404. The absence of this correlation in AD brain suggests that, at least during moderate to late stages of AD (Braak IV–VI), underlying pathogenic mechanisms may disrupt the association between fyn and tau phosphorylated at S202, S262, and S396/404. Notably, inhibition of casein kinase 1 delta, which phosphorylates tau [42], in primary cortical neurons, results in dephosphorylation of tau at multiple sites, including S202, and increases the association of tau with fyn-SH3 [6]. In contrast, increasing tau phosphorylation using the protein phosphatase inhibitor okadaic acid decreases tau binding to fyn-SH3 [6]. An aspartate phosphomimic of tau at S202 has altered affinity for fyn in vitro, dependent on the presence of the microtubule-binding repeats of tau [16]. Phosphorylation at S262 of tau has been shown to result in detachment of tau from microtubules and decreased microtubule stability, as well as inhibition of tau aggregation [42, 43]. Our results could indicate therefore that disassociation of tau from microtubules might affect fyn expression, or that the presence of fyn could influence tau phosphorylation at these residues when tau is not bound to microtubules. It is plausible that tau phosphorylated at S202 and S262 can bind to fyn or influence fyn expression or function under basal conditions, but at late stages of AD, tau becomes increasingly phosphorylated at other residues which could alter the its subcellular localisation or conformation and cause the loss of tau-fyn binding. The progressive accumulation of tau into aggregates in AD could also result in the loss of the association with fyn. Additionally, it was recently shown that tau phosphorylation at S396 is induced following NMDA-receptor mediated long-term depression [4]. Indeed, NMDA receptor activation increased the association of tau phosphorylated at S396/404 with fyn [4]. Taken together, the significant correlation of phosphorylated S396/404 tau with fyn in control brain tissue supports the idea that NMDA receptor activation enhances tau-fyn association. Conversely, the lack of significant correlation between tau phosphorylated at this epitope and fyn in AD brain suggests that pathogenic disease-related mechanisms could interfere with the relationship between phosphorylated tau and fyn. Additional studies involving analysis of fyn kinase activity would likely provide more insight as to the role of fyn in regulating downstream effects of tau phosphorylation and vice versa. Hence, additional investigation is required to fully understand the relationship between fyn and tau phosphorylation, in particular at residues S202, S262, and S396/404 in AD brain.
In this study, identification of the binding site between tau and fyn-SH3 may facilitate the development of compounds that can inhibit tau-fyn interactions, which presents an alternative therapeutic strategy for AD. We also provide evidence that a physiological correlation between phosphorylated tau at S202, S262, and S396/404 and fyn is not present in AD brain, suggesting that progression of disease pathogenesis can influence the relationship between tau and fyn. Further investigation as to the relationship between fyn kinase activity and tau phosphorylation will be important in developing novel therapeutics for AD.
Chinese hamster ovary
CNS:
EDTA:
ethylenediaminetetraacetic acid
glutathione-S-transferase
IPTG:
isopropyl-1-thio-β-D-galactopyranoside
NMDA:
N-methyl-D-aspartate
NSE:
neuron-specific enolase
PSD-95:
post-synaptic density protein 95
SDS:
sodium dodecyl sulphate
SH:
src homology
WT:
wild-type
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We thank Professor Peter Davies (Albert Einstein College of Medicine, New York) for the generous gift of PHF-1, CP27, and CP13 antibodies. Human post-mortem brain tissue was obtained from the London Neurodegenerative Diseases Brain Bank, a member of the Brains for Dementia Research Network.
This work was supported by Alzheimer's Research UK, the Croucher Foundation, BBSRC and Eli Lilly.
Department of Basic and Clinical Neuroscience, King's College London, Maurice Wohl Clinical Neuroscience Institute (K1.24), 125 Coldharbour Lane, London, SE5 9NU, UK
Dawn H. W. Lau, Marte Hogseth, Emma C. Phillips, Amy M. Pooler, Wendy Noble & Diane P. Hanger
Eli Lilly and Company Limited, Lilly Research Centre, Erl Wood Manor, Windlesham, Surrey, GU20 6PH, UK
Michael J. O'Neill
Present address: Nestlé Institute of Health Sciences, Lausanne, Switzerland
Amy M. Pooler
Dawn H. W. Lau
Marte Hogseth
Emma C. Phillips
Wendy Noble
Diane P. Hanger
Correspondence to Diane P. Hanger.
MJO is an employee of Eli Lilly and Company Limited, UK. AMP is now an employee of Nestlé Institute of Health Sciences, Switzerland.
DHWL and DPH designed the experiments and wrote the paper. DHWL performed the majority of the experiments. MH performed the western blots of human post-mortem brain tissue. ECP prepared some of human brain samples. DPH, WN, MJO and AMP supervised the study. All authors read and approved the manuscript.
High intensity scans of western blots of control and Alzheimer's disease (AD) post-mortem human brain tissue from Fig. 4 reveal weak detection of tau phosphorylated at (a) S202 (CP13) and (b) S396/S404 (PHF-1) in control brain tissue, whereas the signal in AD brain is overexposed when shown at this intensity. The molecular weight marker (50 kDa) is shown on the left. (PDF 2638 kb)
Lau, D.H.W., Hogseth, M., Phillips, E.C. et al. Critical residues involved in tau binding to fyn: implications for tau phosphorylation in Alzheimer's disease. acta neuropathol commun 4, 49 (2016). https://doi.org/10.1186/s40478-016-0317-4
PXXP motif


This chapter begins by reminding us that we are made in the likeness of God. Likeness means the quality of being alike, the outward appearance of, the image of. We were made to walk in likeness to God, if we so choose too. Next, the remaining of this short chapter discusses mostly genealogy. Genealogy in the Bible is our way if seeing how we are descendants of Jesus and since we all come from the beginning of God's creations, beginning specifically with Adam and Eve, then we are all brothers and sisters. This is important to sink in absorb, especially when you may be tempted to feel any prejudice towards someone, all people form a family of oneness and one blood so there should never be any room for prejudice or hate in your heart toward another person. More importantly, the genealogy also confirms the Bible's promise that the Messiah, Jesus Christ, would be born into the line of Abraham.
Another thing to take note from this chapter is how much longer the life span was in biblical days compared to today.


大坂屋久左衛門は戦国時代の天正5年に生まれ鉱山開発で成功を収める。のちに鉱工業、オランダ貿易などに進出し住友家に次ぐ豪商となる。大坂屋久左衛門は商家大坂屋代々の当主世襲名として、江戸時代から幕末まで九代に渡り270年間続いた。 Born in 1576, which was during the Warring States Period, Osakaya Kuzaemon succeeded in mine development field. Later, he expanded his business field into mining industry and overseas trade with Nether lands, among others, and Osakaya became the most successful merchant family of the period, after Sumitomo. Osakaya took "Kuzaemon" as their hereditary title, and their family business lasted for 270 years, throughout the Edo Period, followed by 8 successors.


text:
Q: Converting array elements to hashes I have an array with the following elements:
results = ["test=complete", "test2=passed", "test3=failed", "test4=successful"]

I want to convert this array into a hash with the following keys and values:
results_hash = {"test": "complete", "test2": "passed", "test3": "failed", "test4": "successful"}

How can I do this? Not sure if best to use set, hash, etc.

A: In Ruby 2.6:
results.to_h{|s| s.split("=").then{|k, v| [k.to_sym, v]}}
# => {:test=>"complete", :test2=>"passed", :test3=>"failed", :test4=>"successful"}

A: results = ["test=complete", "test2=passed", "test3=failed", "test4=successful"]
results_hash = results.map{|str| str.split("=") }.to_h
# => {"test"=>"complete", "test2"=>"passed", "test3"=>"failed", "test4"=>"successful"}

Edit: As @sawa comments, the keys should be symbols. This is a way:
results.map{|str| a,b = str.split("="); [a.to_sym, b] }.to_h

A: This is my solution:
array = ['test=passed', 'test=failed']

num_of_elements = array.length

x = array[0].split("")

y = array[1].split("")

array = x + y

x.delete_if {|x| x.match('=')}
y.delete_if {|x| x.match('=')}

test_arr = x[0..3].join()
fail = x[4..9].join()

test_arr_2 = y[0..3].join()

fail_2 = y[4..9].join()

results = {}

results.compare_by_identity

results["#{test_arr}"] = fail
results["#{test_arr_2}"] = fail_2

puts results

redpajama_set_name: RedPajamaStackExchange


Rashid Khan slammed an unbeaten 10-ball 34 and recorded figures of 19/3 as Sunrisers Hyderabad beat Kolkata Knight Riders.
Rashid Khan may have surprised many with his absolute blinder with the bat against Kolkata Knight Riders in the second qualifier but for the Sunrisers Hyderabad camp it was 'not a surprise', said veteran Yusuf Pathan. At a time when 150 looked tough, Rashid came out to bat at No 8 and slammed an unbeaten 10-ball 34 as Hyderabad scored 50 runs in the last three overs to post a total of 174/7 in their 14-run victory in Kolkata on Friday night.
"We were never surprised by his knock. Many a times he scored in sixes and boundaries in the Big Bash and other franchise tournaments across the world. He has contributed with quickfire cameos of 20-30s," Yusuf said at the post-match news conference. Tidy bowling by Kolkata spinners Kuldeep Yadav(2/29) and Sunil Narine (1/24) had stifled the Sunrisers after Dinesh Karthik opted to bowl but Rashid changed the complexion of the game.
"He always had the ability to bat that's why he was promoted by one place. Against Chennai also he hit a couple of sixes. We never doubted his ability," Yusuf said. 2016 champions Sunrisers thus snapped a four-match losing streak in the match that mattered most to book the final berth for a second time.
"This was the most important match. We knew we got the liberty to play an extra match in the playoffs by virtue of finishing at the top," Yusuf said. "We believed in our abilities and won today's important match as a team. The two best sides have reached the final and whosoever plays better will win," he said about the summit clash against Chennai in Mumbai tomorrow.
"Chennai have been playing well throughout the season. Our focus will be on playing well and we will hope to continue with the good work." A three-time IPL winner, Yusuf has been going through a lean patch this season as he scored just 215 runs at an average of 23.88 and a modest strike rate of 122.85.
Yusuf, however, said the team was happy with his form. "Hitting sixes is always not needed. We have to see what the team needs and how to play out 20 overs. Maybe sixes are not coming from my bat, but whatever I have played the team is benefiting from that. I am happy with the way I am batting and my coaching staff and skipper is also pleased," he said.


text:
HomeTagsOp-ed
As Florida plans green light for store owners to shoot looters, gunfights at high noon may become the new American reality
Robert Bridge is an American writer and journalist. He is the author of 'Midnight in the American Empire,' How Corporations and Their Political Servants are Destroying the American Dream.
@Robert_Bridge
FILE PHOTO: A sign in front of residents' homes warn away looters four days after Hurricane Irma September 14, 2017 on Stock Island, Florida © Getty Images / Chip Somodevilla
With tensions soaring amid a spate of police incidents involving African Americans, the state of Florida is considering allowing business owners to shoot looters on site. Is this the law the US needs to end mob violence?
Happily, America's Sunshine State has largely escaped the wanton violence and destruction that has greeted others amid a nationwide wave of protests over police brutality. But Governor Ron DeSantis doesn't seem to be taking any chances, empowering the average citizen to behave like the local sheriff. What could possibly go wrong?
The Republican leader drafted tough 'anti-mob' legislation that would give Florida business owners the right to use force against looters participating in "violent or disorderly assemblies." The proposal highlights the deep chasm that now separates Republicans and Democrats over the question of personal security and gun rights at a time when plans to defund police departments are on the drawing board.
Florida Gov. DeSantis' anti-mob law moves to allow citizens to shoot looters https://t.co/snb9IhkOGkpic.twitter.com/VbRXqWaH3B
— Conservative News (@BIZPACReview) November 11, 2020
The nation got a shock in May after George Floyd, a 46-year-old African-American, died under the knee of a white cop during an attempted arrest. The ensuing outbreak of arson, vandalism and looting across the country cost an estimated $1-2 billion, representing the most expensive price tag from civil strife in US history. Meanwhile, many looted and torched small businesses, already on the verge of bankruptcy due to an economic lockdown from Covid-19, were forever consigned to the graveyard.
To put a face on the tragedy, consider the story of Linda Carpenter and her son Scott who fought back tears as they surveyed the charred remains of B & L Office Furniture, a family business they had spent the last 40 years building. In late August, their store in Kenosha, Wisconsin was one of many looted and burned down after local police shot Jacob Blake, a 29-year-old African-American who reportedly resisted arrest. The Carpenters were not guilty of any crime, yet overnight a lifetime of work and dedication to their business was senselessly destroyed.
Although many businesses have 'mob insurance' to cover any damages sustained by looting, theft and vandalism, these same businesses typically do not qualify for coverage from a pandemic. Thus, being forced to rebuild a business in a climate of tremendous economic uncertainty will seem like mission impossible for many. Thus, it may actually come down to a question of sheer survival – keeping food on the table for a family – to protect one's business from literal destruction.
So. If I'm reading this correctly, under DeSantis anti-mob legislation, when conservatives block a Planned Parenthood Clinic, the blockers can legally just be gunned down in Florida. I really don't think they have thought this one through. 🙄
— MC (@mchayden100) November 11, 2020
Not so long ago, Americans could rest assured that the police would protect them and their businesses in times of crisis. Today, no American can take their personal security for granted, and the massive spike in gun sales in the last year is a reflection of that uncertainty. Gun-loving Republicans have been watching in alarm as liberal-run cities and states enact measures that hamper the police from doing their jobs, at the very same time their neighborhoods are going up in flames.
In September, for example, Portland Mayor Ted Wheeler ordered local police to stop using tear gas at protesters during demonstrations. The order came after the city had experienced 105 straight days of protest, many of them violent in nature. Oregon State Police called Wheeler's decision "reckless and short sighted," and threatened to stop sending troopers to help Portland cops deal with the demonstrations.
At the same time, liberals are displaying camaraderie with the looters and vandals, as witnessed by Hollywood celebrities who happily post the criminals' bail and return them to the streets. One Black Lives Matter organizer compared looting that broke out in Chicago to "reparations" that are "owed" to black people as payment for the slavery years. But perhaps the last straw for Republicans came when Mark and Patricia McCloskey responded to protesters trespassing on their property in an upscale St. Louis neighborhood by brandishing firearms. In a decision that infuriated conservatives, the local court charged the McCloskeys with a felony count for the "unlawful use of a weapon."
Clearly, a precedent is being set, at least in the Democrat-run states, that the average citizen has almost no rights when it comes to protecting himself from criminals. That is not the best message to be sending, especially when these same Democrat states are having serious discussions about defunding and even demobilizing their police departments. And as Joe Biden looks set to be the next president, with Kamala Harris and the progressives in the shadows, these lawless tendencies promise to continue.
Sen. Kamala Harris says if she is elected President, she will give Congress 100 days "to get their act together and have the courage to pass reasonable gun safety laws, and if they fail to do it, then I will take executive action" https://t.co/KFdn5Viqm1#HarrisTownHallpic.twitter.com/Ds8DxObhAY
— CNN (@CNN) April 23, 2019
At first glance, Florida's proposal to let business owners resort to deadly force against would-be looters and arsonists seems a bit over-the-top. After all, is it justifiable for any person to lose their life for the crime of stealing a widescreen TV or a pair of sneakers? Governor DeSantis is probably hoping that his legislation will work more as a deterrent to crime than creating dangerous conditions for his people. In other words, if members of the mob understand that business owners are within their legal rights to protect their premises with a gun, then they may think twice before carrying out their illicit activities.
No! That's why we have police. I hate to think I would kill someone over a can of beans. Looting a store is different than robbing a house. Also would they have to be armed?
— Allie Jean (@alliejeanmean) November 11, 2020
The ideal solution, of course, would be for the police to handle such situations as they arise. However, given that these servants of the public are rarely on the scene of a crime when it happens (otherwise it probably wouldn't happen in the first place), who better to protect a business from theft and destruction than the owner himself? So long as the owner is a responsible gun owner, his or her mere presence may go far toward deterring crime. It is even possible to imagine a 'confederation' of armed store owners working together to protect their businesses in times of crisis – something like an armed version of the 'Guardian Angels' – that would serve as a powerful message to any would-be criminals looking for some vulnerable store to loot and torch.
It also does not require much imagination, however, for that scenario to end in disaster as one side begins to seek retribution for the death or injury of one of its own. A bit like mafia groups, nobody would want to leave their home without being armed to the teeth and in a group. In other words, the conditions would be set for the US to return to a 'Wild West' scenario where people openly carried firearms and resolved issues by whoever had the 'fastest draw'. Probably not the most comforting image for a modern democracy, but it is getting difficult to imagine any other.
In the final analysis, it looks as though the liberals have swung too hard left, alienating entire police departments for the crimes of a few. At the same time, the Republicans seem to be overcorrecting the situation, enabling business owners to behave as judge, jury and executioner for petty theft. Neither one of these solutions sounds favorable; best for the American people to place their trust in their police, the majority of whom have the best intentions at heart, while making sure these servants of the people are properly trained to handle any and all situations that may occur on the job.
Otherwise, America is truly heading back to the future of the Wild West.
FloridaUS
Surprising potential Covid-19 remedy: Russian scientists discover mushroom with anti-viral properties
Too tough a punishment? Twitter reacts to New Yorker magazine sacking Jeffrey Toobin after Zoom masturbation fiasco

redpajama_set_name: RedPajamaCommonCrawl


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Transformation Action Planner
The Health Equity Roadmap
A national initiative to drive improvement in health care outcomes, health equity, diversity and inclusion.
Transformation Action Planners (TAPs) are tools to guide AHA members who have completed the Health Equity Transformation Assessment (HETA) to move forward on the continuum of each health equity lever. You need to be logged in as an AHA Member to access the TAPs. The TAPs can also be accessed from the Health Equity Action Library (HEAL).
Systematic and Shared Accountability: Transforming Transformation Action Planner
Action Planning is a critical step in collectively advancing equity and dismantling structural barriers (e.g., racism, sexism, ageism, etc.) in hospitals and health care systems. This Transformation Action Planner is for hospitals at Transforming on the Systematic and Shared Accountability…
Human Resource (HR) Audit
Systematic and Shared Accountability: Affirming Transformation Action Planner
Action Planning is a critical step in collectively advancing equity and dismantling structural barriers (e.g., racism, sexism, ageism, etc.) in hospitals and health care systems. This Transformation Action Planner is for hospitals at Affirming on the Systematic and Shared Accountability continuum.
Systematic and Shared Accountability: Immersing Transformation Action Planner
Action Planning is a critical step in collectively advancing equity and dismantling structural barriers (e.g., racism, sexism, ageism, etc.) in hospitals and health care systems. This Transformation Action Planner is for hospitals at Immersing on the Systematic and Shared Accountability continuum.
Systematic and Shared Accountability: Committing Transformation Action Planner
Action Planning is a critical step in collectively advancing equity and dismantling structural barriers (e.g., racism, sexism, ageism, etc.) in hospitals and health care systems. This Transformation Action Planner is for hospitals at Committing on the Systematic and Shared Accountability continuum.
Systematic and Shared Accountability: Exploring Transformation Action Planner
Action Planning is a critical step in collectively advancing equity and dismantling structural barriers (e.g., racism, sexism, ageism, etc.) in hospitals and health care systems. This Transformation Action Planner is for hospitals at Exploring on the Systematic and Shared Accountability continuum.
Community Collaboration for Solutions: Transforming Transformation Action Planner
Action Planning is a critical step in collectively advancing equity and dismantling structural barriers (e.g., racism, sexism, ageism, etc.) in hospitals and health care systems. This Transformation Action Planner is for hospitals at Transforming on the Community Collaboration for Solutions…
Community Collaboration for Solutions: Affirming Transformation Action Planner
Action Planning is a critical step in collectively advancing equity and dismantling structural barriers (e.g., racism, sexism, ageism, etc.) in hospitals and health care systems. This Transformation Action Planner is for hospitals at Affirming on the Community Collaboration for Solutions continuum.
Community Collaboration for Solutions: Immersing Transformation Action Planner
Action Planning is a critical step in collectively advancing equity and dismantling structural barriers (e.g., racism, sexism, ageism, etc.) in hospitals and health care systems. This Transformation Action Planner is for hospitals at Immersing on the Community Collaboration for Solutions continuum.
Community Collaboration for Solutions: Committing Transformation Action Planner
Action Planning is a critical step in collectively advancing equity and dismantling structural barriers (e.g., racism, sexism, ageism, etc.) in hospitals and health care systems. This Transformation Action Planner is for hospitals at Committing on the Community Collaboration for Solutions continuum…
Community Collaboration for Solutions: Exploring Transformation Action Planner
Action Planning is a critical step in collectively advancing equity and dismantling structural barriers (e.g., racism, sexism, ageism, etc.) in hospitals and health care systems. This Transformation Action Planner is for hospitals at Exploring on the Community Collaboration for Solutions continuum.
Subscribe to Transformation Action Planner


The cornerstone of every new online presence is a website and nowadays it is easy enough to set one up. A designer or developer will be needed to come up with the look and structure of the website. Simple websites may not even need an entirely custom design and could use various templates that are widely available.
Once a website is up and running it will be necessary to promote it so that people actually end up finding it. That particular task will call for the services of an Internet Marketing or online advertising expert who can help coordinate the promotion of the website.
The services of an SEO expert may be necessary if it is decided that the Business wants its website to rank well on search engines and get traffic through that channel. Sometimes the web designer may work with the SEO expert in the planning stage, but that isn't always necessary.
When an offline business shifts online, it exposes itself to a market that spans the globe. If that business is already accustomed to dealing with orders from other countries and already ships worldwide then this isn't a big issue, but if it isn't then it may need to manage the logistics of doing so.
Last but not least, if the business is taking orders online then they will need a payment processing service that can cope with that. Some payment gateways such as PayPal are commonly used and will allow the business to process payments from credit cards in various currencies.
The user can then choose whether to keep the balance in that particular currency or convert it to their normal currency. Sometimes having a balance in other currencies is a good idea – particularly for trading binary options or foreign exchange purposes.
Generally speaking, these are the 5 different options of services that can help a previously offline-only business establish a foothold online. Of course, it is always best to have an overall coordinator who is familiar with the various necessities of the online market while also fluent in the business itself.
Nowadays services such as the ones listed above are abundant and so it should be easy enough to outsource the bulk of the work. In fact, there is really no reason why any business shouldn't have an online presence nowadays considering the advantages that they could gain by doing so, coupled with the fact that with the help of such services it really isn't that hard to get started.
This article is written by Akshay. He is from GetPRed.com and an avid blogger and the author of this post. He has been using the Binary Options Brokers for a smarter and efficient way of making profits. If you wish to write for HBB, kindly check this.
GetPRed provides Guest Posting Service from High Domain Authority Blogs and High Page Rank Blogs making marketing simpler and Affordable.
Should I Start An Online Business?
Great info, but how do you go about finding these experts?
Definitely we need professional technicians at some tasks and we shouldn't ignore it.


The American Council of Learned Societies (ACLS) invites applications in the 2018-19 competition year of The Robert H. N. Ho Family Foundation Program in Buddhist Studies.
In 1865, British polymath Francis Galton published his initial thoughts about the scientific field that would become 'eugenics.' The same year, Russian physician Vasilii Florinskii addressed similar issues in a sizeable treatise, entitled Human Perfection and Degeneration.
The PCB-AHA serves all members of the American Historical Association residing and working in the twenty-two states west of the Mississippi and the four western provinces of Canada. Our annual meeting brings together historians of all spatial, temporal, and thematic specializations.
The Starr Center for the Study of the American Experience invites applications for its full-time residential writing fellowship, which supports outstanding writing on American history and culture by both scholars and nonacademic authors.


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Home/Science/The Blood Moon will seem on Election Day over the past complete lunar eclipse in 3 years
The Blood Moon will seem on Election Day over the past complete lunar eclipse in 3 years
adminNovember 8, 2022
The usually brilliant and glowing moon will seem eerily purple early Tuesday over the past complete lunar eclipse for the following three years. The so-called "Beaver Moon," because the November full moon is thought, will attain its peak brightness at 6:02 a.m. EST through the lunar eclipse.
NASA stated a complete lunar eclipse — when the Solar, Earth and Moon align in order that the Moon passes into Earth's shadow — will happen on Nov. 8, Election Day. The eclipse will start at 3:02 a.m. ET, and the entire eclipse — when the moon is within the darkest a part of the Earth's shadow and seems brilliant purple, incomes it the nickname "blood moon" — will final from about 5:17 a.m. to six: 42 a.m. Jap.
The blood moon part of the eclipse will probably be seen from North and Central America, Ecuador, Colombia, and western elements of Venezuela and Peru. Individuals in Hawaii will have the ability to see each stage of the eclipse, NASA stated.
Map displaying the place the lunar eclipse is seen on November 8, 2022. Contours mark the sting of the seen space throughout contact with the eclipse. The middle of the map is positioned at 168°57′ W, sublunar longitude at mid-eclipse.
NASA Science Imaging Studio
In a video posted on Twitter, NASA stated the eclipse will present a little bit of "sky magic."
"You can see the complete eclipse earlier than dawn, climate allowing, when the moon emerges from the darkish a part of Earth's shadow, known as the umbra," NASA stated.
No particular tools is required to view the eclipse, though being in an space away from brilliant lights will make it extra seen. These with binoculars useful could have the added bonus of seeing the icy planet Uranus, "only a finger's breadth from the eclipsing moon," NASA stated.
There are various causes to look at the sky in November: a complete lunar eclipse, the Leonid meteors and the possibility to see Mars, Saturn and the star Spica! Take a look at the perfect instances to look at these celestial our bodies: pic.twitter.com/hmYGKio4Mq
— NASA (@NASA) November 2, 2022
The final complete lunar eclipse was in Might. This eclipse created what is named "tremendous flower blood moon", which solely happens throughout a complete lunar eclipse, as the complete moon is at its closest level to Earth. Hundreds of thousands have been in a position to spot the large purple area object from the Americas, Europe and Africa.
Lee Cohen is a social media producer and standard reporter for CBS Information who focuses on social justice points.
#Blood #Moon #Election #Day #complete #lunar #eclipse #years
Blood Day Eclipse election lunar moon Total Years
Photo voltaic storm rips gap in Earth's magnetosphere, inflicting extraordinarily uncommon pink auroras
NASA is contemplating SpaceX as an emergency return choice for stranded ISS crew
The NASA mission will present an unprecedented view of the Earth's floor waters | NASA
A suspected meteor lit up the skies of Michigan and past Thursday night time
Analysis exhibits that Earth's oxygen got here from an unexpectedly deep and sizzling supply: ScienceAlert


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redpajama_set_name: RedPajamaGithub


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American Optometric Foundation, Essilor Of America Announce 2011 Technology Grant Recipients At Acad
Rockville, MD – (December 2, 2011) –The American Optometric Foundation (AOF) along with Essilor of America, Inc., the leading manufacturer of ophthalmic lenses, has announced the 2011 recipients of the Essilor Technology Grants; now in its tenth year. The announcement was made at the Foundation's Annual Celebration Luncheon on October 15th during Academy Boston. Three $20,000 grants were awarded to optometry institutions for projects intended to elevate the standards and visibility of ophthalmic dispensing to students and patients in the training environment.
The 2011 recipients are:
State University of New York (SUNY) College of Optometry, New York, NY
University of California Berkeley School of Optometry, Berkeley, CA
The Ohio State University College of Optometry, Columbus, OH
AOF President, Catherine Amos, commented "we are delighted to have a productive ongoing relationship with Essilor. We share their goals of enhancing optometric education and improving patient care."
The American Optometric Foundation is committed to improving the visual welfare of the public by fostering advances in education and research. A leading organization in optometric education, the AOF provides assistance and encouragement to students who plan on pursuing a career in visual health and promotes healthy relationships within and among organizations that share a common goal.
"Essilor is proud to support the next generation of eyecare professionals through its long-standing partnership with the AOF," said Rod Tahran, O.D., F.A.A.O., vice president of professional relations and clinical affairs for Essilor of America, who presented the awards at the luncheon "Since the program began ten years ago, these grants have enabled schools of optometry to make significant advancements their educational programs, ultimately benefitting the entire vision community"
About AOF
The American Optometric Foundation is a philanthropic organization devoted to the advancement of optometric education and research, administering over $400,000 per year in scholarships, fellowships, and research grants. The AOF is an affiliate of the American Academy of Optometry.
About Essilor
Essilor is the leading manufacturer of optical lenses in the United States and is the market leader in progressive, high-index and anti-reflective coated lenses. A pioneer in the development and production of ophthalmic lenses, Essilor employs more than 9,000 people throughout North America. Essilor manufactures optical lenses under the Varilux®, Crizal®, Xperio®, DEFINITY®, Thin&Lite® and other Essilor brand names. Essilor Laboratories of America (ELOA) is the largest, and most trusted, optical lab network in the U.S. and offers a wide choice of services and lens brands, including Essilor premium lenses, to eyecare professionals across the nation. Essilor of America, Inc. (Essilor) is a subsidiary of Paris-based Essilor International, a publicly held company traded on the Euronext Paris stock exchange (Reuters: ESSI.PA).
Varilux, Crizal, Xperio and DEFINITY are registered trademarks of Essilor International. Thin&Lite is a registered trademark of Essilor of America, Inc.


Surprise! 5 Pictures Taken by a Japanese Digital Microscope Camera!
I have found a photo blog on interesting Japanese digital cameras.
Let me introduce a part of the blog.
This blogger has purchased a digital microscope camera called "Anyty".
so he took all kinds of things out of the house and took pictures with the camera.
This is a Japanese 10 yen (about 10 cents) coin.
The coin is enlarged 10 times bigger in this picture.
The engraved Byodo-in temple has a lot of scratches.
It looks totally different. It could have more value than just 10 yen!
Let's take a look at this picture.
The leaf of the apple is like this.
It is finely carved like this. Amazing!
But you find more amazing aspect in natural products.
This is a coffee bean. It looks like a cave.
And, this looks like a sea anemone, doesn't it?
Actually, it is a scrubber.
And, this is a scrubber.
The last one is peach skin.
As you know, peach skin has a smooth touch like fuzzy hair.
But it looks like some kind of a weapon when it is enlarged!
Is Flirting Popular among Married Japanese?


One insightful research piece on the subject is a decade-old piece from Accenture. While we now know more about various shades and details, the broad lines still remain the same: the world has not changed in a fundamental way since 2007, or even since 1998.
The Accenture report quotes a 1998 Corporate Strategy Board study that found that only 17 percent of Fortune 50 companies that experienced a radical slump in revenue growth managed to return back to high or even moderate growth. The conclusion: it is better to start changing when you are still strong, not when you are already weak.
Of course, this is where it all becomes so difficult. Companies do not achieve their peak results when they are at the top of their game. There is considerable inertia between actions and results, and peak results come at a point when the actual performance is already going down.
But how can you tell? Peak result is not a good indicator, because it is a lagging indicator. You can only tell that you have reached the peak after it is already behind you. Furthermore, the implication of the research is that once you can determine the you are past your peak, your odds of getting back in shape are already coming crashing down.
I have personal experience from working with two market-leading companies that came to experience significant issues. What is encouraging is that in both cases, all the signs were there years before any issues arose. What is discouraging is that in both cases, the issues were caused by incompetent management that lasted for years, even close to a decade before it outweighed the legacy of awesomeness that was at the core of both companies.
This all makes perfect sense, of course. Do you believe that it is the poor performance of assembly workers or engineers that causes a company to fail? Sure, overall poor performance contributes to failure, but that poor performance, in turn, is caused by the strategy, management structures, processes, and culture of the company, and those can be guided only by top management. Top management cannot create them at will, but they are the only ones who can change them, with the help of everyone else.
It is not that all management teams are bad. It might even be the case that you can achieve sustainable success with all the same people. It is just a different thing to achieve success once, and to sustain that success over a long period of time. It is easy to fall into complacency, or maybe just not recognize the importance of constant change and stay put when others are moving forwards. Sustaining success requires continuous improvement and the mental strength to keep pushing on even when you are already at the top.
The important realization is that you need to keep acting even if you do not feel the pain. If you fully embrace this view, you can make everything fall into place, even if it takes a mental struggle to force yourself to do it.
Are you continuously improving your way of working? Even those processes that are looking good right now? Do you give your employees time to develop their work, and require them to do so?
Do you know what is going on in your industry and in the world? Do you balance between internal promotions and outside hires? Do you look for a variety of backgrounds when hiring to keep new ideas coming in?
Do you coach your leaders to encourage experimentation? Do you give people autonomy and responsibility?
There are lots of terms to describe doing such things. One could talk about strategic Lean, improvement Kata, coaching Kata, red teaming, and building dynamic capabilities. Many improvement methodologies overlap, so I'm not too interested in tagging things by methodology.
In the end, it is all about acting even when you do not feel the pain, and empowering everyone else in the organization to act as well.


Location of Paysandu on a map.
Paysandu is a city found in Paysandu, Uruguay. It is located -32.32 latitude and -58.08 longitude and it is situated at elevation 49 meters above sea level.
Paysandu has a population of 73,249 making it the biggest city in Paysandu. It operates on the MMT time zone.


MCKINNEY, Texas, Sept. 1, 2015 /PRNewswire/ -- In collaboration with the U.S. Navy, Raytheon Company (NYSE: RTN) recently completed Effective Isotropic Radiated Power (EIRP) testing for its Next Generation Jammer (NGJ) array prototypes at the Benefield Anechoic Facility at Edwards Air Force Base, California.
"Raytheon and the Navy developed a realistic testing program designed to ensure that the NGJ electronic warfare system meets its 2021 initial operating capability commitment," said Travis Slocumb, vice president of Electronic Warfare Systems at Raytheon's Space and Airborne Systems business. "Completion of EIRP testing, while an early milestone, confirms our progress to date and that the program is successfully executing to both schedule and plan."
The prototype testing, conducted over a six week period, indicated that the NGJ will fulfill the U.S. Navy's stringent requirements for EIRP, a prime indicator of the system's range and capacity for reaching and affecting multiple targets simultaneously.
The NGJ is built on a combination of high-powered, agile, beam-jamming techniques and cutting-edge solid-state electronics to achieve two goals: meet the U.S. Navy's electronic warfare mission requirements and provide a cost-effective open systems architecture for future upgrades. It is scheduled to replace legacy ALQ-99 tactical jamming pods, delivering new capabilities for the Navy's EA-18G Growler.


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Home | Publication & Product Detail
Jails in Indian Country, 1998 and 1999
Paula M. Ditton, Bureau of Justice Statistics
July 9, 2000 NCJ 173410
Presents findings from the 1998 and 1999 Surveys of Jails in Indian Country, a complete enumeration of all 69 confinement facilities operated by tribal authorities or the Bureau of Indian Affairs (BIA). The Bureau of Justice Statistics (BJS), in collaboration with the BIA, and the Corrections Program Office and the American Indian and Alaska Native Desk within the Office of Justice Programs, conducted the survey on June 30, as part of BJS' Annual Survey of Jails. The survey includes the number of inmates, staffing, and facility characteristics and needs. The report presents data for each facility, including who runs it, facility age, facility function, rated capacity, authority to house juveniles, number of juveniles held, number of admissions and discharges in last 30 days, inmate deaths, the largest number of inmates held in last 30 days, number of inmates held by gender and conviction status on June 30, number of facility staff by sex and function, facility crowding, renovation and building plans, types of programs available to inmates, and overview of facility and staffing needs.
48 facilities operated by tribal authorities, 20 operated by the BIA, and 1 privately operated facility were supervising offenders in Indian country. The 69 facilities had the capacity to hold 2,118 offenders.
Indian country facilities held 1,621 inmates in custody, and supervised 72 offenders in the community on June 30, 1999. The number of offenders under supervision increased by 8% from the previous year.
In a 1-month period, June 1-30, 1999, facilities in Indian country admitted 8,147 offenders, and discharged 7,744 offenders.
Part of the Jails in Indian Country Series
Full report (PDF 750K)
ASCII file (31K)
Spreadsheet (Zip format 119K)
Codebooks and Datasets
Help for using BJS products
About the Source Data
Survey of Jails in Indian Country (SJIC)
To cite this product, use the following link:
http://www.bjs.gov/index.cfm?ty=pbdetail&iid=1015
View All Publications and Products


The technology used to detect and identify objects has evolved quite a bit over the last several years, and is being used in self-driving cars for recognizing pedestrians, extract data from video, or giving machines the ability to recognize faces and know how you feel. There seems like there's no limit to how this type of artificially intelligent computer vision might be used, and now a team from the University of Western Australia and the International Centre for Radio Astronomy Research (ICRAR) is utilizing that same deep learning technology to identify faraway galaxies in deep reaches of outer space.
Nicknamed ClaRAN (short for Classifying Radio sources Automatically with Neural networks), the algorithm has been adapted from the same facial recognition technology used by Facebook and Microsoft. But instead of scanning faces, ClaRAN scans images from radio telescopes in order to detect so-called radio galaxies — a type of active galaxy nuclei (AGNs) that emit intense jets of light, but at radio wavelengths, rather than within the visible spectrum. And it's from the supermassive black holes at the center of these galaxies that these jets of radio waves emerge.
According to the team, the conventional computer algorithms that are currently being used can identify about 90 percent of these sources of radio emissions.
The team's approach of using AI to cross-match these different data sources seems to work quite well, with a rate of over 90 percent accuracy in pinpointing radio galaxies using radio and infrared images. As the team notes, as our ability to peer deeper and farther into the vastness of the universe improves with better hardware, the amount of data gathered in the next generation of radio surveys (such as the upcoming Evolutionary Map of the Universe) will be enormous — meaning that tools like ClaRAN will be vital in making such research actually feasible in the future.


Published 5/24/2017 . Last updated 9/4/2018 .
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Baking in a HomeAway Condo in Washington, D.C.
Published 11/30/2011 . Last updated 12/21/2018 .


Mixer Sales asked Erich Stauffer to help them get better search engine rankings for their primary keywords, best mixers and mixer sales – and now Mixer Sales is ranking on the first page of Google for 'mixer sales'!
While Mixer Sales ranks #6 for the keyword 'mixer sales', they currently don't rank in the top 100 for 'best mixers', which means we still have some work to do, but we believe in celebrating the small victories. We also helped them setup what we call a 'sticky page' which is a page that stays on top of WordPress blog post listings. This is named after 'sticky posts', but uses a page instead of a post to prevent double posts on the front page. Going forward we're going to recommend on product marketing sites like this a "start with sticky" policy because it provides a kind of launching pad for all of the other posts, which can sometimes get lost. Users can still find posts using categories or tags, but for the most important posts, they should be linked to from the sticky page and possibly even through the top navigation bar. Most of the time we will start off with allowing WordPress to automatically display pages, but as the site ages and becomes more concrete, we'll go back and manually manipulate the page navigation to add posts and other keywords for SEO reasons.
Overall we see good things for Mixer Sales and wish them well in their endeavor to help people find the best deals on kitchen stand mixers, which have become more and more popular over the last several years.
Megamind (voiced by Will Ferrell) is a comedy movie for children ages 7 and older about an evil maniac who finally defeats his long-time nemesis Metro Man (voiced by Brad Pitt) and then merrily goes about laying ruin to Metro City, reveling in the fact that he can now have anything he wants. But then, Megamind's glee quickly turns to dissatisfaction as he realizes that the battle was half the fun and everything now comes too easy. Enter the nerdy Hal (voiced by Jonah Hill) into Tighten, a new hero for Megamind to battle. What Megamind doesn't count on is that Tighten may not turn out to be as good as he's meant to be. Nor does he anticipate falling in love with his old rival's girl Roxanne Ritchi (voiced by Tina Fey). In the end, Megamind finds he must rethink his assumptions about good, bad, and what makes him happy.
Disney's Tangled is a music-filled adventure full of romance and comedy for kids and children ages 6 and older. The movie focuses on Rapunzel, a girl with long magical hair who's lived her entire life imprisoned in a tower by her mother. Naturally optimistic and acquiescent, Rapunzel (voiced by Mandy Moore) rarely complains about her circumstances, but for her 18th birthday she longs to leave the tower to see the floating lights that appear every year on her birthday. Her mother (voiced by Donna Murphy) refuses her request, but when thief Flynn Ryder (voiced by Zachary Levi) climbs the tower to escape his pursuers, Rapunzel eventually decides to trust the young man and convinces him to help her escape to see the floating lights. Tangled is a masterful blend of comedy, adventure, passion, and drama combined with a fantastic music and the type of animation we've come to expect from Disney. The 3D effects add to the experience but most likely won't really be missed in other formats where they are missing.
Did you know there are Android apps made just for the city of Indianapolis and it's attractions?
Search hotels, restaurants, attractions and events based on your location and interests You can also search using Augmented Reality for a street view of what's close by.
Plan your travels by adding preferred destinations to your favorites and generating customized maps.
Redeem cost-saving coupons directly from your phone at top Indianapolis restaurants and attractions.


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Ярун () — озеро в Загребі, у південно-західній частині міського району Трешнєвка-Південь, та прилегла місцевість, названа на честь озера.

Історія
Початки озера походять з того часу, коли там протікала річка Сава. Поселення Ярун згадувалося дуже давно: він позначений на топографічній карті Загреба, датованій 1782 роком. Тоді у ньому було лише дві вулиці — вулиця Ярун і Ярунська набережна.

У ті часи Ярун був рукавом Сави, навколо якого було невеличке поселення і кукурудзяні поля. Повінь 1964 року посприяла будівництву насипу вздовж Сави, який фізично відокремив Ярун від самої річки, перетворивши його на плавні. Тоді з Яруна видобували гравій, який використовували для побудови насипу, тому з часом він поглибився і став озером.

Озеро
Ярун знаходиться на південному краю однойменної місцевості. Його називають також Загребським морем. Тут можна купатися, кататися на водних лижах, ходити під вітрилом, займатися серфінгом; є добре обладнані пляжі з піщаним, гальковим або трав'яним покриттям, де можна приймати сонячні ванни, грати в спортивні ігри, а у пізні години — розважитися в одному з численних нічних клубів або кав'ярень. До озера з центру столиці курсують трамваї маршрутів № 5 і 17.

Озеро та довколишня місцевість — улюблена зона відпочинку жителів Загреба як у спекотні літні дні, так і в холодні зимові. Ярун приваблює охочих до активних видів відпочинку (плавання, біг підтюпцем, катання на роликах і скейтборді, їзда на велосипеді тощо). Окрім розважальних заходів, на озері регулярно проводяться регати з веслування та вітрильного спорту, змагання на байдарках і каное, плавальні марафони, легкоатлетичні виступи, велопробіги тощо. Навколо озера Ярун є «Алея слави» з фотографіями та біографічними відомостями про видатних діячів хорватського спорту.

Крім веслувального каналу завдовжки 2 км, озеро Ярун складається з двох сполучених озер — Великого і Малого та включає 6 островів: Трешнєвка (з'єднаний мостом), Острів Універсіади '87 (з'єднаний мостом), Острів дикої природи, Острів хорватської молоді (з'єднаний мостом), Острів кохання, Острів веслярів (з'єднаний мостом).

Місцевість
Поселення Ярун, підпорядковане однойменному місцевому комітету, охоплює 398,06 га і налічує 12 149 мешканців (2011).

Забудова Яруна багатоквартирними будинками почалася наприкінці 1970-х років, до того Ярун був простим селом. Подальший розвиток пов'язаний із побудовою в 1987 році центру спорту і дозвілля і завдячував проведеній того ж року у Загребі Універсіаді. Нині це мікрорайон середнього класу з дуже низьким рівнем злочинності.

Межі Яруна на півночі утворює вулиця Хорвачанська, яка відділяє його від сусіднього мікрорайону Стаглище, на заході — вулиця Хорватського Сокола і місцевість Врбани, на сході — струмок Врапчак і поселення Гредице, а на півдні — річка Сава. У широкому розумінні Ярун також включає загребські місцевості Стаглище і Гредице.

Примітки

Посилання

 Jarun online
 TD Jarun
 Aquarius Zagreb

Озера Хорватії
Географія Загреба

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text:
Arthur Conan Doyle: The Best Works
Sprache - Englisch
Von Arthur Conan Doyle, Golden Deer Classics
This ebook compiles Conan Doyle's greatest writings, including novels and short stories such as "A Study in Scarlet", "The Hound of the Baskervilles", "The Lost World", "The Adventure of the Speckled Band", "The Sign of Four" and "The White Company".
This edition has been professionally formatted and contains several tables of contents. The first table of contents (at the very beginning of the ebook) lists the titles of all novels included in this volume. By clicking on one of those titles you will be redirected to the beginning of that work, where you'll find a new TOC that lists all the chapters and sub-chapters of that specific work.
Verlag: Oregan Publishing
Genre: Sprache - Englisch
Größe: 1,5 MB
Veröffentlichung: 22. Januar 2018